中国中药杂志

2018, v.43(05) 1008-1013

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荭草花提取物对H_2O_2诱导的HUVEC细胞氧化损伤的保护作用研究
Protective effect of Polygonum orientale flower extract on H_2O_2-induced oxidative damage of HUVEC cells

吴琼;刘香香;陆定艳;李月婷;孙佳;兰燕宇;刘亭;
WU Qiong;LIU Xiang-xiang;LU Ding-yan;LI Yue-ting;SUN Jia;LAN Yan-yu;LIU Ting;Engineering Research Center for the Development and Application of Ethnic Medicine and Traditonal Chinese Medicine,Ministry of Education,Guizhou Medical University;College of Pharmacy,Guizhou Medical University;Provincial Key Laboratory of Pharmaceutics in Guizhou Province,Guizhou Medical University;

摘要(Abstract):

该文首先建立了人脐静脉内皮细胞(HUVEC)过氧化氢(H_2O_2)氧化损伤模型,并筛选了荭草花提取物的低、中、高给药剂量来评价荭草花提取物对H_2O_2所致HUVEC细胞氧化损伤的保护作用并探讨其机制。采用MTS法检测HUVEC细胞活力;生化试剂盒检测细胞乳酸脱氢酶(LDH)漏出量,细胞内丙二醛(MDA)含量及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活力;qRT-PCR和Western blot法分别测定Bcl-2,Bax的mRNA及蛋白水平;Western blot法测定cleaved caspase-3的蛋白水平。结果表明,120μmol·L~(-1)H_2O_2作用0.5 h后,HUVEC细胞存活率下降至55%左右,并且使HUVEC细胞的LDH漏出量、MDA含量增加,SOD及CAT活性降低。此外,H_2O_2还能上调HUVEC细胞中促凋亡蛋白cleaved caspase-3蛋白水平和Bax的mRNA及蛋白水平,下调凋亡抑制蛋白Bcl-2的mRNA及蛋白水平。与H_2O_2模型组相比,50~200 mg·L~(-1)荭草花提取物预处理24 h能明显增加氧化损伤后细胞存活率,降低LDH漏出量、MDA含量,提高SOD及CAT活性,下调促凋亡蛋白cleaved caspase-3蛋白水平和Bax的mRNA及蛋白水平,并上调凋亡抑制蛋白Bcl-2的mRNA及蛋白水平。实验结果提示,荭草花提取物对H_2O_2所致的HUVEC细胞氧化损伤具有一定的保护作用,其机制可能与增强细胞清除自由基能力及抑制细胞凋亡有关。
To investigate the protective effects and mechanism of Polygonum orientale flower extract on H_2O_2-induced oxidative damage of human umbilical vein endothelial cells(HUVEC),H_2O_2 was used to induce the oxidativestress damage on HUVEC cells and efforts were made to screen the low,medium and high drug concentrations of P. orientale flower extract. Cell viability was detected by the MTS assay. The content of lactate dehydrogenase(LDH) and malondialdehyde(MDA),and the activities of superoxidedimutase(SOD) and catalase(CAT) were detected by biochemical kits. The mRNA and protein levels of Bax,Bcl-2 were detected respectively by quantitative real time polymerase chain reaction(qRT-PCR) and Western blot. The protein level of cleaved caspase-3 was detected by Western blot. According to the results,the viability of HUVEC cells was reduced to around 55% after being treated with 120 μmol·L~(-1) H_2O_2 for 0. 5 h. Treatment of H_2O_2 also could increase LDH leakage rate and MDA content and attenuate the activities of SOD and CAT,up-regulate the expression level of Bax and cleaved caspase-3,and down-regulate the expression level of Bcl-2.As compared with H_2O_2 model group,P. orientale flower extract of 50-200 mg·L~(-1) could increase the viability of HUVEC cells,reduce LDH release and MDA content,enhance the activities of SOD and CAT,down-regulate pro-apoptotic protein cleaved caspase-3 and Bax,and up-regulate apoptosis inhibitory protein Bcl-2. In summary,P. orientale flower extract showed a protective effect on H_2O_2-induced HUVEC cells injury,which may result from enhancing the cell capability of clearing the oxygen free radial,decreasing the production of lipid peroxidation and inhibiting apoptosis.

关键词(KeyWords): 荭草花提取物;氧化损伤;细胞凋亡;HUVEC细胞
Polygonum orientale flower extract;oxidative stress injury;apoptosis;HUVEC cells

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81560630,81760699,81360664);; 贵州省科学技术厅人才团队项目(黔科合平台人才[2016]5613/5677)

作者(Author): 吴琼;刘香香;陆定艳;李月婷;孙佳;兰燕宇;刘亭;
WU Qiong;LIU Xiang-xiang;LU Ding-yan;LI Yue-ting;SUN Jia;LAN Yan-yu;LIU Ting;Engineering Research Center for the Development and Application of Ethnic Medicine and Traditonal Chinese Medicine,Ministry of Education,Guizhou Medical University;College of Pharmacy,Guizhou Medical University;Provincial Key Laboratory of Pharmaceutics in Guizhou Province,Guizhou Medical University;

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